Name: testing protozoacidal activity of ligand-lytic peptides against termite gut protozoa in vitro (protozoa culture) and in vivo (microinjection into termite hindgut)
Uploaded: 2010-12-29T17:30:00
Description: Watch this Scientific Journal Video about Testing Protozoacidal Activity of Ligand-lytic Peptides Against Termite Gut Protozoa in vitro Protozoa Culture and in vivo Microinjection into Termite Hindgut

Мы благодарим доктор Элисон Ричард, бывший директор объекта ЛГУ пептид для флуоресцентной синтез лиганда, Interdisciplinaray Центра биотехнологических исследований, UF для лиганд-литического синтеза пептидов, а объект Socolovsky микроскоп для обеспечения доступа к флуоресцентных микроскопов. Финансирование было предоставлено развития SERDP поисковые программы (SEED) из Министерства обороны, Министерства энергетики и охране окружающей среды, биотехнологии AgCenter Междисциплинарная команда программы и штата Луизиана.

Эксперимент 1: Добыча термитов кишечнике простейших в анаэробных условиях <ol><li> Используйте блок вентилятора (Кой Laboratories) в перчаточном ящике постоянно циркулировать воздух через осушитель и тип D катализатором Stak-Пакш для контроля влажности и кислорода и устранения неравномерности ?...

<ol>
	<li>Hansel, W., Leuschner, C., Enright, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Conjugates+of+lytic+peptides+and+LHRH+or+&#946;CG+target+and+cause+necrosis+of+prostate+cancers+and+metastases.">Conjugates of lytic peptides and LHRH or &#946;CG target and cause necrosis of prostate cancers and metastases.</a> Mol. Cell. Endocrinol. 269, 26-33 (2007).</li><li>Husseneder, C., Collier, R. E., Bourtzis, K., Miller, T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Paratransgenesis+for+termite+control.">Paratransgenesis for termite control.</a> Insect Symbiosis. 3, 361-376 (2009).</li><li>Husseneder, C., Grace, J. K., Oishi, D. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+of+genetically+engineered+bacteria+(Escherichia+coli)+to+monitor+ingestion,+loss+and+transfer+of+bacteria+in+termites.">Use of genetically engineered bacteria (Escherichia coli) to monitor ingestion, loss and transfer of bacteria in termites.</a> Curr. Microbiol. 50, 119-123 (2005).</li><li>Husseneder, C., Grace, J. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetically+engineered+termite+gut+bacteria+deliver+and+spread+foreign+genes+in+termite+colonies.">Genetically engineered termite gut bacteria deliver and spread foreign genes in termite colonies.</a> Appl. Microbiol. Biotechnol. 68, 360-367 (2005).</li><li>Javadpour, M. M., Juban, M. M., Lo, W. C., Bishop, S. M., Alberty, J. B., Cowell, S. M., Becker, C. L., Mc Laughlin, M. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=De+novo+antimicrobial+peptides+with+low+mammalian+cell+toxicity.">De novo antimicrobial peptides with low mammalian cell toxicity.</a> J. Med. Chem. 39, 3107-3113 (1996).</li><li>Lai, P. Y., Tamashiro, M., Fuji, J. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Abundance+and+distribution+of+the+three+species+of+symbiotic+protozoa+in+the+hindgut+of+Coptotermes+formosanus+(Isoptera).">Abundance and distribution of the three species of symbiotic protozoa in the hindgut of Coptotermes formosanus (Isoptera).</a> Proc. Haw. Entomol. Soc. 24, 271-276 (1983).</li><li>Leuschner, C., Hansel, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Membrane+disrupting+lytic+peptides+for+cancer+treatments.">Membrane disrupting lytic peptides for cancer treatments.</a> Curr. Pharm. Des. 10, 2299-2310 (2004).</li><li>Mutwiri, G. K., Henk, W. G., Enright, F. M., Corbeil, L. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+the+Antimicrobial+Peptide,+d-Hecate,+on+Trichomonads.">Effect of the Antimicrobial Peptide, d-Hecate, on Trichomonads.</a> J. Parasitol. 86, 1355-1359 (2000).</li><li>Trager, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+cultivation+of+a+cellulose-digesting+flagellate,+Trichomonas+termopsidis,+and+of+certain+other+termite+protozoa.">The cultivation of a cellulose-digesting flagellate, Trichomonas termopsidis, and of certain other termite protozoa.</a> Biol. Bull. 66, 182-190 (1934).</li></ol>

Лиганд-литических пептидов, которые были успешно использованы для эффективного прицельное действие способствует разрушению раковых клеток (Гензель и Лейшнер 2004, Гензель и соавт. 2007). Основываясь на этой концепции, мы разработали гептапептида лиганд, который связывается с поверхн...

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<td>Sigmacote</td>
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<td>Sigma Aldrich</td>
<td>SL-2</td>
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<td>EDANS</td>
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<td>Novabiochem</td>
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<td>Anaerobic glove box</td>
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<td>Coy Laboratories, Inc.</td>
<td>Custom made</td>
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<td>Intellus environmental controller</td>
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<td>Percival</td>
<td>I36NL</td>
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<td>PC-10 Glass micropipette puller</td>
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<td>Narishige Scientific Instrument Lab</td>
<td>PC-10</td>
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<td>Glass needles (Model GD-1, 1 X 900 mm)</td>
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<td>Narishige Scientific Instrument Lab</td>
<td>GD-1</td>
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<td>Leitz micromanipulators</td>
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<td>Vermont Optechs, Inc.</td>
<td>ACS01</td>
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<td>Microinjector</td>
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<td>Tritech Research, Inc.</td>
<td>MINJ-1</td>
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<td>Microcaps</td>
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<td>Drummond Scientific Company</td>
<td>1-000-0005</td>
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<td>LEICA fluorescence imaging system</td>
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<td>Leica</td>
<td>DMRxA2</td>
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<td>LEICA dissecting scope</td>
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<td>Leica</td>
<td>MZ16</td>
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<td>LEICA microscope</td>
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<td>Leica</td>
<td>DMLB</td>
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<td>Olympus dissecting scope</td>
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<td>Olympus</td>
<td>SZ61</td>
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testing protozoacidal activity of ligand-lytic peptides against termite gut protozoa in vitro (protozoa culture) and in vivo (microinjection into termite hindgut)

We are developing a novel approach to subterranean termite control that would lead to reduced reliance on the use of chemical pesticides. Subterranean termites are dependent on protozoa in the hindguts of workers to efficiently digest wood. Lytic peptides have been shown to kill a variety of protozoan parasites (Mutwiri et al. 2000) and also protozoa in the gut of the Formosan subterranean termite, Coptotermes formosanus (Husseneder and Collier 2009). Lytic peptides are part of the nonspecific immune system of eukaryotes, and destroy the membranes of microorganisms (Leuschner and Hansel 2004). Most lytic peptides are not likely to harm higher eukaryotes, because they do not affect the electrically neutral cholesterol-containing cell membranes of higher eukaryotes (Javadpour et al. 1996). Lytic peptide action can be targeted to specific cell types by the addition of a ligand. For example, Hansel et al. (2007) reported that lytic peptides conjugated with cancer cell membrane receptor ligands could be used to destroy breast cancer cells, while lytic peptides alone or conjugated with non-specific peptides were not effective. Lytic peptides also have been conjugated to human hormones that bind to receptors on tumor cells for targeted destruction of prostate and testicular cancer cells (Leuschner and Hansel 2004).
In this article we present techniques used to demonstrate the protozoacidal activity of a lytic peptide (Hecate) coupled to a heptapeptide ligand that binds to the surface membrane of protozoa from the gut of the Formosan subterranean termite. These techniques include extirpation of the gut from termite workers, anaerobic culture of gut protozoa (Pseudotrichonympha grassii, Holomastigotoides hartmanni, Spirotrichonympha leidyi), microscopic confirmation that the ligand marked with a fluorescent dye binds to the termite gut protozoa and other free-living protozoa but not to bacteria or gut tissue. We also demonstrate that the same ligand coupled to a lytic peptide efficiently kills termite gut protozoa in vitro (protozoa culture) and in vivo (microinjection into hindgut of workers), but is less bacteriacidal than the lytic peptide alone. The loss of protozoa leads to the death of the termites in less than two weeks.
In the future, we will genetically engineer microorganisms that can survive in the termite hindgut and spread through a termite colony as "Trojan Horses" to express ligand-lytic peptides that would kill the protozoa in the termite gut and subsequently kill the termites in the colony. Ligand-lytic peptides also could be useful for drug development against protozoan parasites.

Watch this Scientific Journal Video about Testing Protozoacidal Activity of Ligand-lytic Peptides Against Termite Gut Protozoa in vitro Protozoa Culture and in vivo Microinjection into Termite Hindgut

Testing Protozoacidal Activity of Ligand-lytic Peptides Against Termite Gut Protozoa in vitro Protozoa Culture and in vivo Microinjection into Termite Hindgut

Мы существующему порядку демонстрации того, что лиганды связываются с поверхностью мембраны целлюлозы переваривания простейших в кишечнике Формозы подземных термитов помощью флуоресцентной микроскопии и лигандов, что в сочетании с литических пептидов убивать этих простейших<em&gt; В пробирке</em&gt; (Анаэробных простейших культуры) и<em&gt; В естественных условиях</em&gt; (Инъекции в кишке термитов).

Тестирование Protozoacidal активность лиганд-литических пептиды против термитов Gut Protozoa В пробирке (Protozoa культуры) и В естественных условиях (Микроинъекция в Термит кишке)

We are developing a novel approach to subterranean termite control that would lead to reduced reliance on the use of chemical pesticides. Subterranean ...

Research

JoVE Journal

Biology

Testing Protozoacidal Activity of Ligand-lytic Peptides Against Termite Gut Protozoa in vitro (Protozoa Culture) and in vivo (Microinjection into Termite Hindgut)

Extracting DNA from the Gut Microbes of the Termite (Zootermopsis Angusticollis) and Visualizing Gut Microbes

Extracting DNA from the Gut Microbes of the Termite Zootermopsis Angusticollis and Visualizing Gut Microbes

Извлечение ДНК из Gut микробы Термит (Zootermopsis Angusticollis) и Визуализация Gut Микробы

Termites are among the few animals known to have the capacity to subsist solely by consuming wood.  The termite gut tract contains a dense and ...

Investigating the Microbial Community in the Termite Hindgut - Interview

Исследование микробного сообщества в задней кишке термитов - Интервью

Jared Leadbetter explains why the termite-gut microbial community is an excellent system for studying the complex interactions between microbes.     The ...

Layers of Symbiosis - Visualizing the Termite Hindgut Microbial Community

Слои Симбиоз - Визуализация Микробная Термит кишке сообщества

Jared Leadbetter takes us for a nature walk through the diversity of life resident in the termite hindgut - a microenvironment containing 250 different ...

Testing the Physiological Barriers to Viral Transmission in Aphids Using Microinjection

Тестирование физиологические барьеры на пути передачи вируса в Тли Использование Микроинъекция

Potato loafroll virus (PLRV), from the family Luteoviridae infects solanaceous plants. It is transmitted by aphids, primarily, the green peach aphid. When ...

In vivo and in vitro Studies of Adaptor-clathrin Interaction

In vivo and in vitro Studies of Adaptor-clathrin Interaction

В естественных условиях и в пробирке исследования Адаптер-клатрин Взаимодействие

A major endocytic pathway initiates with the formation of clathrin-coated vesicles (CCVs) that transport cargo from the cell surface to endosomes1-6. CCVs ...

Activation of Apoptosis by Cytoplasmic Microinjection of Cytochrome c

Activation of Apoptosis by Cytoplasmic Microinjection of Cytochrome c

Активация апоптоза Цитоплазматическая Микроинъекция цитохрома С</em

Apoptosis, or programmed cell death, is a conserved and highly regulated pathway by which cells die1.  Apoptosis can be triggered when cells encounter a ...

Induction and Testing of Hypoxia in Cell Culture

Индукция и тестирование Гипоксия в культуре клеток

Hypoxia is defined as the reduction or lack of oxygen in organs, tissues, or cells.  This decrease of oxygen tension can be due to a reduced supply in ...

Dissection of Xenopus laevis Neural Crest for in vitro Explant Culture or in vivo Transplantation

Dissection of Xenopus laevis Neural Crest for in vitro Explant Culture or in vivo Transplantation

Рассечение<em&gt; Xenopus Laevis</em&gt; Нервного гребня для<em&gt; В пробирке</em&gt; Эксплантата Культура или<em&gt; В естественных условиях</em&gt; Трансплантация

The neural crest (NC) is a transient dorsal neural tube cell population that undergoes an epithelium-to-mesenchyme transition (EMT) at the end of ...

In Vivo Microinjection and Electroporation of Mouse Testis

In Vivo Microinjection and Electroporation of Mouse Testis

В Vivo микроинъекции и электропорации мыши яичка

This video and article contribution gives a comprehensive description of microinjection and electroporation of mouse testis in vivo. This particular ...

Functional Reconstitution and Channel Activity Measurements of Purified Wildtype and Mutant CFTR Protein

Функциональная Реорганизация и активности канала измерения очищенной дикого типа и мутантных CFTR белка

The Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) is a unique channel-forming member of the ATP Binding Cassette (ABC) superfamily of ...